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EDITORIAL FOCUS
1Department of Physiology and Biophysics, University of Southern California Keck School of Medicine, Los Angeles, California; and 2The Water and Salt Research Center, Department of Cell Biology, Institute of Anatomy, University of Aarhus, Aarhus, Denmark
Submitted 10 February 2007 ; accepted in final form 9 May 2007
The distal convoluted tubule (DCT) Na+-Cl– cotransporter (NCC), the target of thiazide diuretics, is responsible for the reabsorption of 5–10% of filtered NaCl. The aim of this study was to test the hypothesis that acute infusion of the angiotensin-converting enzyme (ACE) inhibitor captopril (at 12 µg/min) for 20 min provokes trafficking of NCC from apical plasma membranes (APM) to subapical cytoplasmic vesicles (SCV), which is reversed by acute ANG II infusion (ANG II at 20 ng·kg–1·min–1 along with 12 µg/min captopril) for 20 min in male Sprague-Dawley rats (250–350 g). By immuno-electron microscopy using an anti-NCC (D. Ellison) 71.5 ± SD 4.9% of the NCC gold labeling was associated with the APM in control, sham operated, and infused rats, while captopril infusion reduced NCC in APM to 54.9 ± 6.9% (P < 0.001) and markedly increased immunogold labeling of SCV. Subsequent infusion of ANG II with captopril restored NCC immunogold labeling of APM to 72.4 ± 4.2%, that is, 20% of the total NCC trafficked between APM and SCV. Likewise, on density gradients of cortex, captopril provoked redistribution of 27.3% of total NCC from low-density APM-enriched membranes to higher-density membranes and ANG II+captopril restored 20.3% of the NCC to APM-enriched fractions. Redistribution occurred independent of a change in NCC total abundance. In conclusion, this study demonstrates that ACE inhibition provokes acute trafficking of NCC out of the plasma membrane, which likely decreases DCT Na+ reabsorption, while ANG II provokes rapid trafficking of NCC from stores in subapical vesicles to the plasma membrane, which likely increases DCT Na+ reabsorption.
sodium transport; thiazide receptor; immunoelectron microscopy
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